KR102240357B1 - Antioxidant composition for cosmetics containing a compound isolated from extract of Rosa rugosa Thumb - Google Patents

Abstract

The present invention relates to an antioxidant composition for cosmetics, characterized in that it contains as an active ingredient at least one compound selected from the group consisting of compounds of Formulas 1 to 6 isolated from the maemaehwa extract.
Antioxidant composition for cosmetics containing the compound isolated from the maemaebu extract as an active ingredient effectively separates the compounds 1 to 6 from the ethyl acetate fraction of the maebubu extract and can be usefully used as a cosmetic composition having an antioxidant component. There is.

Description

Antioxidant composition for cosmetics containing a compound isolated from extract of Rosa rugosa Thumb}

The present invention relates to a cosmetic antioxidant composition containing as an active ingredient a compound isolated from a maebu extract, and more particularly, a fractionated extract obtained by extracting maebu with a solvent and then using water and ethyl acetate as fractionation solvents, respectively. It relates to a composition having an excellent antioxidant effect containing a compound separated by as an active ingredient.

Plum blossom refers to the flower buds of the flower belonging to the Rosaceae family. This flower is a deciduous broad-leaved shrub that grows naturally in the sandy area of the beach or at the foot of the mountain. The leaves are dislocated, the small leaves are oval or obovate, the bottom is sharp, the ends are blunt or round, serrated, and there are preoccupants and villi on the back side. The flower is red and blooms in May, and the fruit is the most globular, yellow-red, and ripens in August. Chemical components of glycosylation include β-sitosterol, campesterol, euscaphic acid, β-sitosteryl-β-D-glucopyanoside, campesteryl-β-D-glucopyranoside, rosamultin, arjunetin, and (+)-catechin.

Maedanghwa is a drug that enters the nose of the nose and liver, allows blood to pass through, benefits Gi, and cures wind. As an anti-inflammatory drug, it is used for blood circulation, landscaping, stomach pain, Yuongjongtong, menstrual discomfort, rheumatism, and bruises.

In recent years, the antioxidant activity of the extracts and fractions of barberry has been recognized, but there has been no report on the antioxidant activity of specific compounds.

KR 10-2013-0065185 A (2013. 06. 19.) KR 10-1825179 B1 (2018.01.29.)

The present invention has been conceived to solve the problems of the prior art, and an object to be solved in the present invention is to provide an antioxidant composition for cosmetics containing a compound isolated from a maebustion extract as an active ingredient.

In addition, it is an object of the present invention to provide a method for separating the compounds represented by Chemical Formulas 1 to 6 from the extract of maebustion.

Cosmetic antioxidant composition containing as an active ingredient the compound isolated from the maebustion extract according to the present invention for achieving the above object is one selected from the group consisting of the compounds of Formulas 1 to 6 isolated from the maebustion extract It is characterized by containing the above compound as an active ingredient.

In addition, the extraction solvent of the plume extract is characterized in that water, alcohol, or a mixture thereof.

In addition, the compounds of Formulas 1 to 6 isolated from the maebu hwa extract is characterized in that it is isolated from the ethyl acetate fraction of the maebu hwa extract.

In addition, the compounds of Formulas 1 to 6 isolated from the lumped extract used at least one selected from the group consisting of column chromatography, high performance liquid chromatography, ion exchange chromatography, affinity chromatography, and size exclusion chromatography. It is characterized by separating.

In addition, it is characterized in that it comprises 0.1 to 10% by weight of the separated compound as an active ingredient from the ethyl acetate fraction of the lumped extract.

Cosmetic antioxidant composition containing as an active ingredient the compound isolated from the maebustion extract according to the present invention is effectively used as a cosmetic composition having an antioxidant component by separating compounds 1 to 6 from the ethylacetate fraction of the maebustion extract efficiently. There is an effect that can be.

1 is a view showing the separation process of compounds 1 to 6 separated from the mass formation according to the present invention.
Figure 2 is a diagram showing the DPPH activity of the ethyl acetate fraction of the maekkaethwa extract according to the present invention.
Figure 3 is a diagram showing the DPPH activity of the 50% methanol fraction according to the present invention.
Figure 4 shows the DPPH activity for the six compounds isolated from the 50% methanol fraction according to the present invention.
Figure 5 is a view showing the results of UPLC analysis of the six compounds isolated from the extract according to the present invention.
Figure 6 is a view showing the ultraviolet spectrum of the six compounds isolated from the extract according to the present invention.
7 is a view showing the UPLC-HR-ESI-MS spectrum of the six compounds isolated from the extract according to the present invention.
Figure 8 is a diagram showing the 1H NMR spectrum of Compound 1 of the six compounds isolated from the extract according to the present invention.
Figure 8 is a diagram showing the 1H NMR spectrum of Compound 1 of the six compounds isolated from the extract according to the present invention.
9 is a view showing the 1H NMR spectrum of Compound 2 of the six compounds isolated from the extract according to the present invention.
FIG. 10 is a diagram showing a 1H NMR spectrum of Compound 3 among 6 compounds isolated from the extract of maegaehwa according to the present invention.
11 is a diagram showing the 1H NMR spectrum of compound 5 among the six compounds isolated from the maemaehwa extract according to the present invention.
12 is a diagram showing the 1H NMR spectrum of compound 6 among the six compounds isolated from the maemaehwa extract according to the present invention.

The present invention relates to an antioxidant composition for cosmetics containing as an active ingredient at least one compound selected from the group consisting of compounds of Formulas 1 to 6 isolated from the extract of maegaehwa.

[Formula 1]

Compound 1 represented by Formula 1 may be referred to as Rutin.

MW (molecular weight): 610.51

Molecular Formula: C ₂₇ H ₃₀ O ₁₆

[Formula 2]

Compound 2 represented by Formula 2 may be referred to as Hyperoside.

MW (molecular weight): 464.37

Molecular Formula: C ₂₁ H ₂₀ O ₁₂

[Formula 3]

Compound 3 represented by Chemical Formula 3 may be referred to as Isoquercitrin.

MW (molecular weight): 464.09

Molecular Formula: C ₂₁ H ₂₀ O ₁₂

[Formula 4]

Compound 4 represented by Chemical Formula 4 may be referred to as Hyperin 6''-gallate.

MW (molecular weight): 616.48

Molecular Formula: C ₂₈ H ₂₄ O ₁₆

[Formula 5]

Compound 5 represented by Chemical Formula 5 may be referred to as Avicularin.

MW (molecular weight): 434.08

Molecular Formula: C ₂₀ H ₁₈ O ₁₁

[Formula 6]

Compound 6 represented by Chemical Formula 6 may be referred to as 2-Phenylethyl 6-O-galloyl-β-D-glucopyranoside.

MW (molecular weight): 436.14

Molecular Formula: C ₂₁ H ₂₄ O ₁₀

The compounds of Formulas 1 to 6 isolated from the maebu hwa extract are isolated from the ethyl acetate fraction of the maebu hwa extract, and the compounds isolated from the maebu hwa extract are five flavonoid compounds and one phenolic compound.

First, the lumping extract is extracted with a hydrophilic solvent, and the hydrophilic solvent may be water, alcohol, or a mixture thereof. The alcohol may be ethanol, methanol, n-propanol, isopropanol, n-butanol, sec-butanol, isobutanol, tert-butanol, or a mixture thereof, but is not limited thereto. In the present invention, the extract is extracted using 100% ethanol as an extraction solvent for the lumping extract.

In addition, in preparing a fraction using a fractionation solvent for the lumped extract, in the present invention, a fraction was prepared using ethyl acetate.

In addition, the compound separated from the ethyl acetate fraction of the massif extract may be chromatography or filtration, and the chromatography may be column chromatography, high performance liquid chromatography (HPLC), ion exchange chromatography, affinity chromatography, and size. It may be exclusion chromatography or a combination thereof.

The compound isolated from the ethylacetate fraction of the maegaehwa extract can be used as a cosmetic composition having an antioxidant component.

In addition, with respect to the total composition, it is preferable to include 0.1 to 10% by weight of the compound isolated from the ethylacetate fraction of the maebustion extract as an active ingredient.

It is preferable that the content of the composition is more than the minimum value so as to achieve the minimum antioxidant effect, and the content of the composition is preferably less than the maximum value in consideration of the reduction in feeling of use and applicability to various formulations due to excessive addition.

Hereinafter, the present invention will be described in more detail through examples. However, the following examples are for illustrative purposes only, and are not intended to limit the scope of the present invention, which is obvious to those skilled in the art. That is, simple modifications or changes of the present invention can be easily implemented by a person skilled in the art, and all such modifications or changes can be considered to be included in the scope of the present invention.

Example 1: Preparation of the extract, fraction and separation of the compound from the fraction

1 is a diagram showing the separation process of compounds 1 to 6 separated from buried formation.

Referring to Figure 1 to describe the method of separating the compound from the fractions and the extract, the fraction and the fractions, 1 kg of the cultivation was extracted with 3L of 100% ethanol at 50°C. 70 g of the extracted barberry ash extract was suspended in 2 L of water and fractionated with ethyl acetate. The ethyl acetate fraction was evaporated under reduced pressure to give 26 g of a brown residue. The solvent fraction was tested for DPPH- scavenging activity. Among them, the ethyl acetate fraction showed antioxidant activity.

26 g of the fractionated fraction was subjected to reverse phase column chromatography (YMC-gel RP-C18, 70) by step gradient elution using methanol and water (0:100, 30:70, 50:50, 70:30 and 100:0). -230 mesh, 500g) was eluted with each 500mL to obtain 5 fractions (Fr. 1 to 5). Again Fr. 3 (12.8 g) was further purified by RP column chromatography using a stepwise gradient of methanol-water (step by step 20:80 to 100:0; 500 mL) to obtain 12 fractions (Fr.3-1 to Fr.3). -12) 3-6 (1.1g) was sequentially separated by HPLC (YMC-Pack ODS-A, 5㎛, φ20 × 150mm, UV 254nm, flow rate 2ml/min), and Compound 1 (16.5mg) , Compound 2 (21.4 mg), Compound 3 (48.1 mg), Compound 4 (4 mg), Compound 5 (6.3 mg) and Compound 6 (40.7 mg) were obtained.

Figure 2 shows the DPPH activity of the ethyl acetate fraction and the fraction of the maehwa extract, Figure 3 shows the DPPH activity of the 50% methanol fraction, Figure 4 is the DPPH activity for the six compounds isolated from the 50% methanol fraction Is shown.

Experimental Example 1: Analysis of 6 compounds isolated from the extract of maebustion

(1) UPLC/MS analysis

The six compounds isolated from the maehwa extract were analyzed by UPLC/Q-TOF MS.

UPLC/MS analysis was performed on a Waters ACQUITY UPLC system combined with a Waters SYNAPT G2-Si high-definition mass spectrometer. Chromatographic separation was performed at 35° C. using an ACQUITY UPLC HSS C18 (100 mm×2.1 mm, 1.8 μm) column, and at 10° C. auto sampler temperature. Mobile phase A consists of water and mobile phase B consists of acetonitrile. The solvent initially changes from 20% B to 20-30% B over 50 minutes at a flow rate of 0.3 mL/min (Table 1). It was then washed for 15 minutes with 95% B before the next sample injection. The chromatogram was monitored at 254 nm using a UV detector.

For UPLC-HR-MS analysis, capillary voltage and cone voltage were set to 2.0 kV in negative ion electrospray mode.

Mass spectrometry was calibrated during acquisition using an external reference (Lock-Spray) consisting of 300 ng/mL solution of leukine enkephalin (Sigma Aldrich) at a flow rate of 20 nL/min, and negative ion mode ([MH]) to ensure accuracy and reproducibility. -m / z = 554.2615) to generate a reference ion. The results are shown in FIG. 5.

(2) HPLC-PDA analysis

HPLC analysis was performed on a Shimadzu-HPLC system (Shimadzu Corporation, Kyoto, Japan) and included a CBM-20A controller, LC-20AP pump and SPD-M20A photodiode array (PDA) detector. Chromatographic separation was achieved on a Luna C18 reverse phase column (250 mm x 4.6 mm, 5 μm). Analysis conditions are shown in Table 2 below.

(3) DPPH radical scavenging ability measurement

DPPH (1,1-diphenyl-2-picrylhydrazyl) analysis is the most widely used method of measuring antioxidant activity, and all experiments were performed three times. Ascorbic acid was used as a positive control, and the antioxidant activity profile was generated by measuring the radical scavenging ability of each fraction using DPPH (1,1-diphenyl-2picrylhydrazyl). Each fraction was dried with a centrifuge using a vacuum concentrator and reconstituted in methanol to make a 10 mg/mL stock solution. All 22 stock solutions were diluted with methanol and mixed with 190 μL of 150 μM DPPH aqueous solution.

The absorbance of the remaining DPPH was measured at 517 nm after 20 minutes as follows.

Ab _t = absorbance of DPPH solution using the tested extract

Ab ₀ = absorbance of DPPH solution upon addition of ethanol

Ab _b = absorbance of the extraction solution tested by adding methanol.

(4) result

The UV spectra were compared with each of the 6 compounds and the 6 compounds isolated from the cultivated extract of the present invention, and the 6 compounds were identified as 5 flavonoid compounds and one phenolic compound. The structure of the compound was established by HR-ESI-MS, NMR and NMR analysis in the literature. The results are shown in FIG. 6.

In addition, the structure of the six compounds isolated from the extract of the present invention was determined by NMR analysis by comparison with chemical evidence, physical methods, and structures reported in the literature. The results are shown in Fig. 7 and Tables 3 and 4 below.

As can be seen from Table 4, when comparing compounds 1 to 6, in the order of (6)> (5)> (3)> (1)> (2)> (4), free radicals for these compounds You can check the antioxidant activity through the curl experiment. Among the six isolated and purified compounds, compound 6 showed the highest antioxidant effect.

Claims (5)

In the cosmetic composition having an antioxidant component,
1 kg of crushed mass was extracted with 3L of 100% ethanol at 50°C, and 70 g of the extracted mass lumped extract was suspended in 2L of water and fractionated with ethyl acetate, and methanol and water were added to the ethyl acetate fraction by weight of 0:100, 30 :70, 50:50, 70:30, and 100:0 were eluted by reverse-phase column chromatography to obtain 5 fractions, and the fraction obtained by dividing methanol and water at a weight ratio of 50:50 among the fractions was methanol. -Twelve fractions were obtained by purification by reverse phase column chromatography using a stepwise gradient of water, and a compound consisting of Formulas 1 to 6 separated through high performance liquid chromatography was contained as an active ingredient,
Cosmetic antioxidant composition comprising 0.1 to 10% by weight of the compound isolated from the ethyl acetate fraction of the maebustion extract as an active ingredient.

[Formula 1]

[Formula 2]

[Formula 3]

[Formula 4]

[Formula 5]

[Formula 6]

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